8 Jun 2011



Primary cell culture
The maintenance of growth of cells dissociated from the parental tissue (such as kidney, liver) using the mechanical or enzymatic methods, in culture standard using apposite schooner or plastic containers is called Primary Cell Culture.
The primary cell culture may possibly be of two types depending in the lead the kind of cells in culture.

A) marina Dependent /Adherent cells- Cells publicized to require attachment pro growth are fit to be marina Dependent cells. The Adherent cells are ordinarily derived from tissues of organs such as kidney everywhere they are immobile and embedded in connective tissue. They grow adhering to the cell culture.

B) Suspension Culture/Anchorage Independent cells - Cells which sort out not require attachment pro growth or sort out not glue to the go up of the culture vessels are anchorage self-determining cells/suspension cells. All suspension cultures are derived from cells of the blood logic since these cells are furthermore floating in plasma in vitro e.G. Lymphocytes.
Secondary cell cultures
When a primary culture is sub-cultured, it becomes renowned as secondary culture or cell line. Subculture (or passage) refers to the conveying of cells from lone culture vessel to a further culture vessel.
Subculturing- Subculturing or splitting cells is vital to periodically provide fresh nutrients and growing interval pro continuously growing cell shape. The process involves removing the growth media, washing the plate, disassociating the adhered cells, ordinarily enzymatically. Such cultures could be called secondary cultures.
Cell Line
A Cell Line or Cell Strain could be restricted or unremitting depending in the lead whether it has restricted culture life span or it is immortal in culture. On the basis of the life span of culture, the cell shape are categorized into two types:
A) Finite cell Lines - The cell shape which be inflicted with a restricted life span and energy through a restricted digit of cell generations (usually 20-80 population doublings) are renowned as Finite cell shape. These cell shape exhibit the property of friend inhibition, density limitation and anchorage dependence. The growth rate is gradual and doubling calculate is around 24-96 hours.
B) Continuous Cell Lines - Cell shape transformed under laboratory conditions or in vitro culture conditions produce increase to unremitting cell shape. The cell shape trade show the property of ploidy (aneupliody or heteroploidy), absence of friend inhibition and anchorage dependence. They grow in monolayer or suspension form. The growth rate is rapid and doubling calculate is 12-24 hours.
C) Monolayer cultures - When the underside of the culture vessel is covered with a unremitting layer of cells, ordinarily lone cell in thickness, they are referred to as monolayer cultures.
D) Suspension cultures - Majority of unremitting cell shape grow as monolayers. Some of the cells which are non-adhesive e.G. Cells of leukemia or particular cells which can be mechanically reserved in suspension, can be propagated in suspension. There are particular advantages in procreation of cells by suspension culture method.
These advantages are:
(a) The process of procreation is much nearer.,
(b) The frequent replacement of the standard is not vital.,
(c) Suspension cultures be inflicted with a fleeting lag cycle,
(d) behavior with trypsin is not vital,
(e) a homogenous suspension of cells is obtained,
(f) the maintenance of suspension cultures is straightforward and bulk production of the cells is straightforwardly achieved.,
(g) scale-up is furthermore very well-located.
The cell shape are renowned by:
A) A code e.G. NHB pro Normal Human intellect.
B) A cell line number- This is applicable as several cell shape are derived from the same cell culture source e.G. NHB1, NHB2.
C) Number of population doublings, the cell line has already undergone e.G. NHB2/2 earnings two doublings.

The cell shape are characterized by their a) growth rate and b) karyotyping.
A) Growth Rate - A growth curve of a fastidious cell line is established taking into consideration the population doubling calculate, a lag calculate, and a infiltration density of a fastidious cell line. A growth curve consist of:
1) Lag Phase: The calculate the cell population takes to recover from such secondary culture, glue to the culture vessel and apply.

2) Log Phase: Inside this period the cell digit begins to boost exponentially.

3) Plateau Phase: During this period, the growth rate slows or stops due to exhaustion of growth standard or confluency.
B) Karyotyping - Karyotyping is valuable as it determines the species of origin and determine the boundary of gross chromosomal changes in the line. The cell shape with abnormal karyotype are furthermore used if they take up again to go habitual function. Karyotype is affected by the growth conditions used, the way in which the cells are subcultured and whether or not the cells are frozen.
There are particular stipulations with the intention of are associated with the cell shape.
These are as follows:
(i) Split ratio- The divisor of the dilution ratio of a cell culture by subculture.
(ii) Passage number- It is the digit of era with the intention of the culture has been educated.,
(iii) Generation number- It refers to the digit of doublings with the intention of a cell population has undergone...

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